Custom antibody development

We offer world-leading antibody development platforms based on a proprietary RabMAb® rabbit monoclonal platform, a phage display platform (AxioMx), and a next-generation sequencing platform (NGS-RabMAb®)

Applications

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application

Abreviews

Notes

IHC-FoFr

1/300.

ICC

Use at an assay dependent concentration.

ICC/IF

1/200.

WB

Use a concentration of 1 µg/ml. Detects a band of approximately 170,45 kDa (predicted molecular weight: 126,39 kDa).Can be blocked with Rat Nogo peptide (ab32297).

IHC-Fr

1/200.

Target

Function

Developmental neurite growth regulatory factor with a role as a negative regulator of axon-axon adhesion and growth, and as a facilitator of neurite branching. Regulates neurite fasciculation, branching and extension in the developing nervous system. Involved in down-regulation of growth, stabilization of wiring and restriction of plasticity in the adult CNS. Regulates the radial migration of cortical neurons via an RTN4R-LINGO1 containing receptor complex (By similarity). Isoform 2 reduces the anti-apoptotic activity of Bcl-xl and Bcl-2. This is likely consecutive to their change in subcellular location, from the mitochondria to the endoplasmic reticulum, after binding and sequestration. Isoform 2 and isoform 3 inhibit BACE1 activity and amyloid precursor protein processing.

Tissue specificity

Isoform 1 is specifically expressed in brain and testis and weakly in heart and skeletal muscle. Isoform 2 is widely expressed except for the liver. Isoform 3 is expressed in brain, skeletal muscle and adipocytes. Isoform 4 is testis-specific.

Sequence similarities

Contains 1 reticulon domain.

Domain

Three regions, residues 59-172, 544-725 and the loop 66 amino acids, between the two transmembrane domains, known as Nogo-66 loop, appear to be responsible for the inhibitory effect on neurite outgrowth and the spreading of neurons. This Nogo-66 loop, mediates also the binding of RTN4 to its receptor.

Cellular localization

Endoplasmic reticulum membrane. Anchored to the membrane of the endoplasmic reticulum through 2 putative transmembrane domains.

Immunofluorescent staining for Nogo in rat brain cerebellum using Rabbit polyclonal to Nogo (ab32298; 1/300 incubated for 18h at RT); immunoreactivity was detected in most brain areas. The staining is quite diffuse on cell bodies, axons and dendrites. The picture was acquired using X20 objective. Protocol details: Rats were intracardially perfused with 4% paraformaldehyde. Whole brain tissue was post-fixed overnight in the same fixative, and cryoprotected in 20% sucrose and frozen in OCT. 30µm coronal sections were cut by cyrostat for use in fre floating IHC. Secondary antibody Alexa fluor 488 1/1000 was incubated for 2 hours at room temperature.

Immunocytochemistry stainning for Nogo in Cor1 neural stem cells from mouse using Rabbit polyclonal to Nogo (ab32298; 1/200 incubated for 1h at RT); Immunoreactivity was detected in cell-body as well as in neurites.

ab32298 staining Nogo in human SH-SY5Y cells by Immunocytochemistry/ Immunofluorescence.Cells were fixed in paraformaldehyde, blocked with 10% serum for 20 minutes at 24°C, then incubated with ab32298 at a 1/200 dilution for 16 hours at 4°C. The secondary used was an Alexa-Fluor 488 conjugated donkey anti-rabbit polyclonal used at a 1/1000 dilution. Counterstained with Hoechst 33258 (blue).

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